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Use of 16S rRNA gene-targeted group-specific primers for real-time PCR analysis of predominant bacteria in human feces.
- Source :
-
Applied and environmental microbiology [Appl Environ Microbiol] 2004 Dec; Vol. 70 (12), pp. 7220-8. - Publication Year :
- 2004
-
Abstract
- 16S rRNA gene-targeted group-specific primers were designed and validated for specific detection and quantification of the Clostridium leptum subgroup and the Atopobium cluster. To monitor the predominant bacteria in human feces by real-time PCR, we used these specific primers together with four sets of group-specific primers for the Clostridium coccoides group, the Bacteroides fragilis group, Bifidobacterium, and Prevotella developed in a previous study (T. Matsuki, K. Watanabe, J. Fujimoto, Y. Miyamoto, T. Takada, K. Matsumoto, H. Oyaizu, and R. Tanaka, Appl. Environ. Microbiol. 68:5445-5451, 2002). Examination of DNA extracted from the feces of 46 healthy adults showed that the C. coccoides group was present in the greatest numbers (log10 10.3 +/- 0.3 cells per g [wet weight] [average +/- standard deviation]), followed by the C. leptum subgroup (log10 9.9 +/- 0.7 cells per g [wet weight]), the B. fragilis group (log10 9.9 +/- 0.3 cells per g [wet weight]), Bifidobacterium (log10 9.4 +/- 0.7 cells per g [wet weight]), and the Atopobium cluster (log10 9.3 +/- 0.7 cells per g [wet weight]). These five bacterial groups were detected in all 46 volunteers. Prevotella was found in only 46% of the subjects at a level of log10 9.7 +/- 0.8 cells per g (wet weight). Examination of changes in the population and the composition of the intestinal flora for six healthy adults over an 8-month period revealed that the composition of the flora of each volunteer remained stable throughout the test period.
- Subjects :
- Adult
Bacteria classification
Bacteria genetics
Bacteroides fragilis genetics
Bacteroides fragilis isolation & purification
Bifidobacterium growth & development
Bifidobacterium isolation & purification
Clostridium genetics
Clostridium isolation & purification
Genes, rRNA
Humans
Prevotella genetics
Prevotella isolation & purification
RNA, Ribosomal, 16S genetics
Species Specificity
Bacteria isolation & purification
DNA Primers
Feces microbiology
Polymerase Chain Reaction methods
Subjects
Details
- Language :
- English
- ISSN :
- 0099-2240
- Volume :
- 70
- Issue :
- 12
- Database :
- MEDLINE
- Journal :
- Applied and environmental microbiology
- Publication Type :
- Academic Journal
- Accession number :
- 15574920
- Full Text :
- https://doi.org/10.1128/AEM.70.12.7220-7228.2004